Scrapie of sheep is the prototype of a group of diseases collectively designated transmissible spongiform encephalopathies (TSE). TSE diseases affect a wide range of species including sheep, cattle, mink, humans, deer, elk and others. The newest member of the group, bovine spongiform encephalopathy (BSE) or mad cow disease is believed to have occurred as the result of cattle being fed bone meal and protein supplements contaminated with tissue derived from scrapie-infected sheep. There is no evidence that sheep scrapie infects humans. However, passage of sheep scrapie through cattle appears to have altered its biological characteristics making BSE transmissible to people. Which TSE diseases will transmit to which other species is difficult to predict but probably involves complex interactions between specific protein molecules of the species considered and in particular a protein designated prion protein (PrP). The overall objective of this project is to determine how PrP and other factors and conditions govern transmission from one species to another. Strategies to interfere with the transmission or disease process will be developed based on these findings. Interspecies transmission of TSE diseases between and among various species is now known to involve interactions between the prion proteins (PrP) of the species involved. For example, the TSE agent which causes disease in mice can in a few circumstances cause disease in hamsters but the hamster agent does not cause clinical disease in mice. To determine how PrP influences transmission from hamsters to mice we have developed transgenic mice which express hamster prion protein (HPrP). The HPrP has been expressed using a variety of promoters including the neuron- specific enolase promoter (NSE) which targets expression to neurons, the GFAP specific promoter to target expression to astrocytes and natural promoters which result in expression in many tissues. Each of these types of mice has been bred to PrP null mice which do not express mouse PrP. Thus, we have available several types of mice which we have used to investigate how expression of PrP in specific cell types influences susceptibility to scrapie and how HPrP and mouse PrP (MoPrP) interact. Mice which express HPrP under the control of natural or NSE promoters are completely susceptible to hamster scrapie agent following intracerebral inoculation. This result showed that PrP is critical to transmission and that expression restricted to neurons is sufficient for transmission to occur. If NSE/HPrP was expressed in mice which lacked functional MoPrP expression, the incubation period was reduced indicating that the presence of MoPrP in some way competed with HPrP to delay the onset of disease. The effect was protective for the mice in that disease was either delayed or completely circumvented. Mice which express HPrP under the control of the GFAP promoter (GFAP/HPrP) did not become clinically sick after inoculation of hamster scrapie agent. However, if GFAP/HPrP was expressed in mice which lacked MoPrP, expression they did become sick following hamster agent inoculation. This result suggested that astrocyte specific expression of HPrP was also sufficient for transmission to occur though much less efficiently than was true if expression was limited to neurons. Furthermore, this result demonstrated very strong interference between MoPrP and HPrP in the mice which expressed both. Because TSE diseases are thought to be transmitted orally we also inoculated the NSE/HPrP Tg mice and another Tg line designated Tg7, where HPrP is expressed in multiple tissues, with hamster scrapie agent orally and intraperitoneally. Most of the mice inoculated by these routes survived if they expressed both mouse and HPrP. These results suggested that therapeutic intervention in TSE diseases could be based on these interference mechanisms. We also found that hamster scrapie agent was able to persist in mice following intracerebral inoculation for the normal lifespan of the mouse and in the abscence of clinical scrapie ( mice had previously been considered totally resistant to hamster scrapie agent). This persistence was dependent on the presence of PrP. We determined the kinetics of hamster agent clearance from mice and showed that although most agent is rapidly eliminated that which persists can eventually adapt to the new species but required blind passage to a second recipient group. Brain and spleen homogenates obtained from the second pass mice caused clinical disease in hamsters as well as mice(3rd pass). On fourth passage various strains had evolved with their own characteristic phenotypes. Thus, hamster agent persisted through at least four serial passages in a species thought to be resistant. This result is consistent with the theory that BSE was derived from sheep but suggests that the disease in cattle was not manifest as clinical disease until more than a primary passage in cattle had occurred. Persistence and eventual adaptation to a resistant species suggests that similar situations could occur in other species combinations. In the USA there is concern that chronic wasting disease of deer and elk could be transmissible to other species including humans though the host range has not yet been fully delineated. We have increased our effort with regard to CWD and have initiated studies to determine more precisely the pathogenesis of CWD including the importance of various tissues, possibilities regarding better diagnostic tests and investigating the potential for transmission to humans.